Influenza virus‐specific T cell‐mediated cytotoxicity: integration of the virus antigen into the target cell membrane is essential for target cell formation

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Abstract
This study deals with the requirements for target cell recognition by influenza A virus‐ specific cytotoxic T lymphocytes (CTL). H‐2‐identical cells were incubated with infectious or UV light‐inactivated influenza A virus expressing either cleaved or uncleaved hemagglutinin (HA). Thereafter, the treated cells were tested in a 4‐h 51lCr assay for susceptibility to CTL‐mediated cytolysis.Regardless whether the influenza virus was infectious, virions expressing cleaved HA were efficient in target cell formation. In contrast, cells incubated with either active or UV‐inactivated virions expressing uncleaved HA were not lysed by virus‐ specific CTL. Yet, after mere trypsin‐mediated cleavage of the HA of cell‐absorbed viroins, strong cytolysis could be observed. On the other hand, solubilization of the envelope lipid bilayer by ethyiether abolished the capacity of the remaining HA to induce target cell formation. The results clearly suggest that mere adsorption of virions to the membrane of cells, which is performed by virus with uncleaved HA, is insufficient for target cell formation. For this, both cleaved HA and an intact envelope appear to be crucial. We conclude that fusion of the virion into the cell membrane is essential for target cell formation.